Termination of Deoxyribonucleic Acid in
            <i>Escherichia coli</i>
            by 2′,3′-Dideoxyadenosine

Toji, Lorraine; Cohen, Seymour S.

doi:10.1128/jb.103.2.323-328.1970

Cited by 27 publications

(1 citation statement)

References 16 publications

(16 reference statements)

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“…If the ddNs ai-e phosphorylated to 2',3'-dideoxynucleoside-5'triphosphates (ddNTPs) they become analogs of the 2'deoxynucleoside-5'-triphosphates (dNTPs) which normally serve as substrates for cellular and viral DNA polymerases. Previous studies have shown that 2',3'dideoxyadenosine triphosphate (ddATP) inhibits E. coli DNA polymerase in vitro (Toji and Cohen, 1970) and that 2',3'-dideoxythymidine-5 '-triphosphate (ddTTP) differentially inhibits eukaryotic cellular and viral DNA polymerases. Cellular DNA polymerase a is relatively resistant to ddTTP while cellular DNA polymerases p and y are relatively sensitive (Edenberg et al, 1978;Waqar et al, 1978;van der Vliet and Kwant, 1978;Abboud and Horwitz, 1979;Ono et al, 1979;Krokan et al, 1979;Wist, 1979;McLennan, 1980;Allaudeen, 1980).…”

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confidence: 99%

Effects of 2′,3′‐dideoxynucleosides on mammalian cells and viruses

Waqar

Evans

Manly

et al. 1984

Journal Cellular Physiology

127

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Previous studies on the biological effects of the 2',3'-dideoxynucleosides (ddNs) have shown that while ddAdo is lethal to E. coli, ddThd has minimal effects on the growth of mammalian cell lines and that it inhibits retrovirus infection of some cell lines but not others. Previous studies have also shown that the 5'-triphosphate of ddThd, ddTTP, selectively inhibits cellular DNA polymerases beta and gamma and retroviral reverse transcriptases. Cellular DNA polymerase alpha is relatively resistant to ddTTP. We have extended these findings to show that the 5'-triphosphates of the other 3 ddNs (ddATP, ddCTP, and ddGTP) affect cellular DNA polymerases alpha, beta, and gamma in the same fashion as does ddTTP. We also show that all four ddNs in concentrations up to 100 microM have negligible effects on the growth of NIH Swiss 3T3 cells. These negligible effects may be due to inefficient intracellular phosphorylation of each nucleoside to the triphosphate. We have determined that, in several different cell lines, ddThd is phosphorylated only at a very slow rate to ddTTP, and in the one cell line tested (monkey CV-1 cells), ddAdo and ddGuo are also poorly phosphorylated. Both ddAdo and ddGuo, and probably ddThd, are converted by CV-1 cells to additional unknown compounds which may have biological activity. The four ddNs display effects of different magnitudes on certain virus infections. Although 30 microM ddThd inhibits herpes simplex I infection of CV-1 cells by 50%, 30 microM ddAdo has no effect. Infection of NIH Swiss 3T3 cells by 334C murine leukemia virus is inhibited 70-80% by ddAdo, ddCyd, and ddThd at 50 microM, but inhibition by 50 microM ddGuo is 100%.

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confidence: 99%

Effects of 2′,3′‐dideoxynucleosides on mammalian cells and viruses

Waqar

Evans

Manly

et al. 1984

Journal Cellular Physiology

127

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The mechanisms of lethal action of arabinosyl cytosine (araC) and arabinosyl adenine (araA)

Cohen

1977

Cancer

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Certain D-arabinosyl nucleosides, notably arabinosyl cytosine (araC) and arabinosyl adenine (araA), are useful in the treatment of certain leukemias and some DNA virus infections, respectively. The compounds are lethal to animal cells and some bacteria. Despite extensive deamination, the parent nucleosides are transported within sensitive cells and phosphorylated to the mono-, di-and triphosphates. AraCTP and araATP are good specific competitive inhibitors of tumor cell or virus-induced DNA polymerases, competing with dCTP and dATP respectively. In addition to markedly inhibiting DNA synthesis, the aranucleotides enter newly formed DNA in internucleotide linkage. Sensitivity to the nucleosides appears to correlate with the relative ratio of formation of the triphosphate via a nucleoside kinase to degradation of the nucleoside via a nucleoside deaminase. Inhibition of the deaminase increases formation of the aranucleoside triphosphate in leukemic or virus-infected cells and markedly increases the toxicity of the nucleosides. Combinations of inhibitors of the deaminases and of the aranucleoside are being explored in clinical situations. In addition, the slow penetration of aranucleotides into cells has been observed and some of these 5'-phosphates are useful antiviral agents, e.g., against herpes virus in herpetic keratitis.Cawm 40:509-518, 1977HIS REVIEW HAS BEEN LIMITED TO A DIS-

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Penetration of mouse fibroblasts by 2′‐deoxyadenosine 5′‐phosphate and incorporation of the nucleotide into DNA

Plunkett¹,

Cohen

1977

Journal Cellular Physiology

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The metabolism of 2'-deoxyadenosine S'-phosphate (dAMP) by exponentially growing mouse fibroblasts (L-cells) in suspension culture has been studied. Cells incubated for four hours with 0.10 pmole [3H,32P]dAMP/ml in medium containing 9.6 pmole 31Pi/ml incorporate both activities at nearly linear rates into acid-soluble and acid-insoluble fractions. The 3H/32P value increased about 30-fold in each fraction during the incubation, indicating extensive dephosphorylation of dAMP. The DNA from treated cells was degraded enzymatically to 5'-mono-nucleotides, which were fractionated by ion-exchange chromatography. 3H was associated exclusively with dAMP (53%) and dGMP (47%). 32P, associated with all deoxynucleotides, was at a 20-fold higher specific activity in dAMP than in either dGMP or dCMP. The specific activity of [ 32P]dAMP incorporated into DNA in four hours was 24-fold greater than that of 32Pi in the cellular pool. In experiments in which cells were incubated with 32Pi plus or minus 0.10 @mole dAMP/ml, the specific activity of dAMP was slightly less than that of dGMP or dCMP, which were equal. These results suggest that the higher specific activity of [32P]dAMP in the DNA of cells after incubation with doubly-labeled dAMP was due to the intact penetration of some dAMP into the cells with its subsequent incorporation into DNA. Calculations, based on the amount of exogenously added [3H]thymidine incorporated into the cellular DNA in parallel cultures and the 32P of dAMP isolated from this DNA, suggest that 1% of the total dAMP residues incorporated during the 4-hour incubation were derived directly from intact, exogenously added dAMP.We have previously reported that upon incubation of L-cells with the toxic nucleotide 9-P-D-arabinosyladenine 5'-phosphate (araAMP), small quantities of the compound entered the cells without prior dephosphorylation (Plunkett et al., '74). After penetration, the original araAMP moiety was phosphorylated to the triphosphate, araATP, and incorporated into DNA.These results provided a biochemical basis for the suggestion that, in certain cases, nucleotides might be more toxic formulations for chemotherapeutic purposes than nucleosides. As nucleotides, the compounds might prevent the degradation of the nucleoside moiety by cellular enzymes or could possibly bypass cellular deficiencies in the enzymes required for activation of the toxic compound. In the former case, we have provided evidence that araAMP and its cyclic derivative, 3', 5'-cyclic araAMP, are more toxic to L-cells than is 9-p-D-arabinofuranosyladenine (araA) (Ortiz et al., '72; Plunkett and Cohen, '75b). In the latter case, we have shown

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Termination of Deoxyribonucleic Acid in Escherichia coli by 2′,3′-Dideoxyadenosine

Cited by 27 publications

References 16 publications

Effects of 2′,3′‐dideoxynucleosides on mammalian cells and viruses

Effects of 2′,3′‐dideoxynucleosides on mammalian cells and viruses

The mechanisms of lethal action of arabinosyl cytosine (araC) and arabinosyl adenine (araA)

Penetration of mouse fibroblasts by 2′‐deoxyadenosine 5′‐phosphate and incorporation of the nucleotide into DNA

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